EXPERIMENTS -6

AIM- (RHS & LHS)

To prepare the coloured components present in the given grass/flower by
ascending paper chromatography and determine their Rf values.

Theory- (RHS)
The basic principle of chromatographic technique is based on the
differential migration of the individual components of a mixture through a
stationary phase under the influence of moving phase. The stationary phase
may be a porous solid (such as slilca, alumina,etc) packed in a column or
supported on a filter paper or a glass strip. The moving (or mobile) phase
may be some solvent or a gas and is referred as an eluent.

Apparatus - (RHS)
Gas jar, glass rod , filter paper strip ( Whatman No.1 filter paper) , jar cover,
fine capillary tube.

Requirement- (RHS)
A mixture of grass/flower , alcohol and distilled water.

Procedure-(RHS)

1. Take a Whatman Filter paper strip (20 x 2) and draw a line with pencil
above 3cm from one end.Draw another line lengthwise from the centre of
the paper.
2. With using capillary tube, put a drop of mixture of grass and flower at the
point P. let it dry in air. Put another drop on the same spot and dry
again.Repeat 2-3 times, so that the spot is rich in mixture.
3. Suspend the paper vertically in a gas jar containing the solvent (eluent) a
way that the pencil line (and the spot) remains about 2cm above the solvent
level (50% alcohol + distilled water).
4. Cover it keep undisturbed. Notice the rising solvent, along with the
mixtures of grass/flower, after the solvent has risen about 15cm you will
notice two different spots of colour on the filter paper.
5. Take the filter paper out of the jar and mark the distance that the solvent
has risen on the paper with a pencil. This is called the solvent front.
6. Dry the paper. Put pencil marks in the centre of the colour spots.
7. Measure the distance of the two spots from the original line and the
distance of the solvent from the original line.
8. Calculate the Rf values of pigments (spots) present in the grass/flower by
using formula.
Rf = Distance travelled by the spot from point of application
Distance travelled by the solvent from original line

OBSERVATIONS AND CALCULATIONS:- (LHS)

SR No Colour travelled by Distance travelled Distance travelled by Rf values

the spot by the spot from the solvent from
original line original line
1.

2.

Result - (LHS)
Rf value of green component is = ____ cm
Rf value of red component is = ____ cm
Rf value of yellow component is = ____ cm

Percautions - (RHS)
1. Always make use of a fine capillary tube.
2. Keep the strip erect. Do not let it to be curled.
3. Do not allow the spot to dip in the solvent.